Antibodies to lipoproteins and apolipoproteins and methods of use thereof

Abstract

Compositions and methods using antibodies which are immunoreactive with specific apolipoproteins to determine the concentrations of lipoproteins such as HDL and LDL, and / or apolipoproteins in human blood, serum or plasma sample, are described. Monoclonal antibodies (MAbs) are described that specifically bind to epitopes present in apolipoproteins and lipoproteins, enabling rapid and reliable determinations of levels of specific blood lipoprotein and / or apolipoprotein levels, including Apo B-100, Apo A-I, Apo A-II, Apo C-III, and Apo E, and thereby determination of relative ratios of HDL and LDL and LpaI and LpaII. In a preferred embodiment, the compositions are strips of a solid phase material coated with one or more of the antibodies and are referred to herein as “dipsticks”. The dipsticks specifically bind a lipoprotein or apolipoprotein when dipped into a protein sample. The amount of lipid associated with a bound lipoprotein or the amount of apolipoprotein bound on the dipstick is quantitated using an appropriate method, for example, by staining with a lipid stain or reaction with a second labelled antibody. The intensity of the stain on the dipstick is proportional to the concentration of the lipoprotein lipid or apolipoprotein circulating in the blood and can be quantitated by comparison with standards containing known amounts of lipid.

Description

[0001]This application is a continuation of U.S. Ser. No. 08 / 268,809 now U.S. Pat. No. 6,107,045 filed Jun. 30, 1994.BACKGROUND OF THE INVENTION[0002]This invention is generally in the field of methods and compositions for the determination and quantitation of lipoproteins and apolipoproteins in human blood.Human Plasma Lipoproteins and Apolipoproteins[0003]Plasma lipoproteins are carriers of lipids from the sites of synthesis and absorption to the sites of storage and / or utilization. Lipoproteins are spherical particles with triglycerides and cholesterol esters in their core and a layer of phospholipids, nonesterified cholesterol and apolipoproteins on the surface. They are categorized into five major classes based on their hydrated density as very large, triglyceride-rich particles known as chylomicrons (less than 0.95 g / ml), very low density lipoproteins (VLDL, 0.95 to 1.006 g / ml), intermediate-density lipoproteins (IDL, 1.006 to 1.019 g / ml), low-density lipoproteins (LDL, 1.019 ...

AI Technical Summary

Benefits of technology

The patent describes a method for using antibodies to determine the levels of specific lipoproteins and apolipoproteins in human blood. These antibodies can be used to create dipsticks that can quickly and reliably determine the levels of HDL, LDL, Apo B-100, Apo A-I, Apo A-II, Apo C-III, and Apo E. The dipsticks can be used without a physician or technical laboratory and can be provided in kits for easy use. The antibodies can also be used in other methods such as enzyme immunoassays, radioimmunoassays, fluorescent and chemiluminescent immunoassays, and purification of the apolipoprotein or lipoprotein with which they are immunoreactive.

Problems solved by technology

Many epidemiological and clinical studies have shown that increased LDL levels in the blood are associated with an increased risk of CHD.

Furthermore, postmenopausal women have a significant increase in CHD risk while their HDL concentrations decrease.

However, the ultracentrifugation methods are too expensive and time consuming for the purpose of measuring LDL-C levels to assess lipoprotein abnormalities and CHD risk in routine clinical application.

The time required for analysis, typically 24 hours, is the major difficulty with agarose column chromatography.

The development of high-performance liquid chromatography (HPLC) methods has reduced the analysis time, but has increased the cost and complexity of the procedure.

However, most investigators have found that the precipitation methods are plagued with systematic errors when samples with high TG levels are analyzed.

Although atherogenic, these lipoprotein particles are not commonly measured in routine clinical laboratories.

Therefore, a low C-III and E ratio is associated with increased risk of CHD because it reflects either high VLDL and IDL and normal HDL or, more frequently, high VLDL and IDL combined with low HDL.